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Development and Validation of a Multiplex Non-HLA Antibody Assay for the Screening of Kidney Transplant Recipients

Elena G Kamburova, Tineke Kardol-Hoefnagel, Bram W Wisse, Irma Joosten, Wil A Allebes, Arnold van der Meer, Luuk B Hilbrands, Marije C Baas, Eric Spierings, Cornelis E Hack, Franka E van Reekum, Arjan D van Zuilen, Marianne C Verhaar, Michiel L Bots, Adriaan C A D Drop, Loes Plaisier, Jan Meeldijk, Niels Bovenschen, Marc A J Seelen, Jan Stephan Sanders, Bouke G Hepkema, Annechien J A Lambeck, Laura B Bungener, Caroline Roozendaal, Marcel G J Tilanus, Christina E Voorter, Lotte Wieten, Elly M van Duijnhoven, Mariƫlle A C J Gelens, Maarten H L Christiaans, Frans J van Ittersum, Shaikh A Nurmohamed, Neubury M Lardy, Wendy Swelsen, Karlijn A M I van der Pant, Neelke C van der Weerd, Ineke J M Ten Berge, Frederike J Bemelman, Paul J M van der Boog, Johan W de Fijter, Michiel G H Betjes, Sebastiaan Heidt, Dave L Roelen, Frans H Claas, Henny G Otten

Published: 01/01/2018

Abstract

The best treatment for patients with end-stage renal disease is kidney transplantation. Although graft survival rates have improved in the last decades, patients still may lose their grafts partly due to the detrimental effects of donor-specific antibodies (DSA) against human leukocyte antigens (HLA) and to a lesser extent also by antibodies directed against non-HLA antigens expressed on the donor endothelium. Assays to detect anti-HLA antibodies are already in use for many years and have been proven useful for transplant risk stratification. Currently, there is a need for assays to additionally detect multiple non-HLA antibodies simultaneously in order to study their clinical relevance in solid organ transplantation. This study describes the development, technical details and validation of a high-throughput multiplex assay for the detection of antibodies against 14 non-HLA antigens coupled directly to MagPlex microspheres or indirectly via a HaloTag. The non-HLA antigens have been selected based on a literature search in patients with kidney disease or following transplantation. Due to the flexibility of the assay, this approach can be used to include alternative antigens and can also be used for screening of other organ transplant recipients, such as heart and lung.

Full Access Link: Frontiers in immunology